转基因大动物常用载体频数分析及筛查策略

摘要

构建重组表达载体是转基因动物生产制备研究中非常关键的一步,包括构建完整的外源基因表达盒,包含目的基因、调控序列(启动子、终止子)和筛选报告基因等.本文概述了转基因大动物制备技术,归纳统计了近10年转基因猪、牛、羊制备过程中常用的载体和频数,统计结果表明,转基因猪中启动子频数从多到少依次为酪蛋白、CAG、CMV启动子,终止子频数依次为兔β-globin polyA、酪蛋白polyA、SV40 polyA和BGH poly A;转基因羊中启动子频数从多到少依次为酪蛋白、BLG和CMV启动子,终止子依次为酪蛋白polyA、BLGpolyA、BGH poly A、SV40 poly A和兔β-globin poly A;转基因牛中启动子频数从多到少依次为酪蛋白、CMV、人乳清白蛋白启动子等,终止子依次为SV40 poly A、BGH poly A和酪蛋白polyA.根据统计结果提出针对启动子和终止子的多重检测和筛查策略;分析了未来转基因动物检测可能存在的问题,以期对转基因动物的监管和筛查检测方法的建立提供参考.%The recombinant expression vector is vital in production of genetically modified animals,which includes complete exogenous gene expression cassette containing the target gene promoter,terminator and reporter gene.This paper briefly introduces the genetically modified animal production technology and counts the vector frequency commonly used in the production of genetically modified pigs,cattle and sheep during nearly 10 years.The statistical results show that promoter frequency of the genetically modified pigs from more to less are casein,CAG,CMV promoters,terminator frequency are rabbit β-globin poly A,casein poly A,SV40 poly A and BGH poly A;The promoter frequency in genetically modified sheep from more to less are casein,BLG and CMV promoters and terminator are casein poly A,BLG poly A,BGH poly A,SV40 poly A and rabbit β-globin poly A;The promoter frequency in genetically modified cattle from more to less are casein,CMV and human α-lactalbumin promoters and terminator are SV40 poly A,BGH poly A and casein poly A.Multiple detection and screening strategies aiming at the promoter and terminator are proposed according to the statistical results.This paper also analyses the possible problems in the future in genetically modified animal detection,this review will provide reference for regulatory perspective and detection of genetically modified animals.